Methods for single nucleotide polymorphism detection

a single nucleotide polymorphism and detection method technology, applied in the direction of specific use bioreactors/fermenters, organic chemistry, after-treatment of biomass, etc., can solve the problems of high cost of arrays, high cost of synthesizing, and troublesome secondary structures

Inactive Publication Date: 2003-10-14
MONOGRAM BIOSCIENCES
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Problems solved by technology

However, neither LSSP-PCR nor SSCP gives specific sequence information and both depend on the questionable assumption that any base that is changed in a sequence will give rise to a conformational change that can be detected.
However both methods have the serious limita

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Embodiment Construction

In accordance with the subject invention, a rapid and accurate method is provided for detecting a plurality of single nucleotide polymorphisms in a nucleic acid sample. The method employs as reagents, a mixture of particles, template dependent polynucleotide polymerase, and at least one chain terminating labeled nucleoside triphosphate. The particles have a primer which hybridizes to a target sequence which has a snp at the 5' end of the target sequence (the primer is 3' of the snp in relation to the sequence to which it hybridizes and the snp is 3' of the primer sequence) and a coding composition, which coding composition has one or more entities which can be determined free of the particle. Each coding composition in the mixture of particles is unique for the primer bound to the same particle to which the coding composition is bound.

The protocols which are employed will depend on the number of snps to be determined from a single sample. There will usually be at least 10 snps of in...

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Abstract

Methods and compositions are provided for determining large numbers of single nucleotide polymorphisms in target DNA employing particles having (1) primers complementary to sequences in the target DNA where the next succeeding 3'-nucleotide is a potential single nucleotide polymorphism and coding composition members, where the members are unique for each primer, and (2) differentially labeled terminating nucleotides, where the label permits separation of the terminating nucleotides. Desirably the particles are separated into groups having a common prevalent next succeeding nucleotide. The particles and target DNA are combined under nucleotide extending conditions, the particles separated into groups in accordance with the terminating nucleotide and the coding members identified, so that one knows the sequence and the single nucleotide polymorphism. Various protocols are provided for the determination.</PTEXT>

Description

1. Field of the InventionThe field of this invention is the detection of the simultaneous detection of large numbers of variations in DNA sequences.2. BackgroundIndividual susceptibility to disease and environmental toxins, prognosis of existing disease, efficacy of a particular drug, susceptibility to adverse drug reactions, etc., in humans and domestic animals and plants are becoming increasingly predictable by genetic analysis. Many of these characteristics are associated with multiple genes and are not due to a single genetic abnormality. The extensive effort currently devoted to genome sequencing has revealed a strong correlation between sequence polymorphism, particularly base deletions and substitutions and the occurrence of numerous genetic diseases. It has been estimated that single nucleotide polymorphisms occur at about 1 in every 400 nucleotides, a frequency that is continually updated as the human genome is unraveled. Single nucleotide polymorphisms therefore provide a ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6827C12Q1/6834C12Q2565/519C12Q2535/125C12Q2563/179C12Q2563/131
Inventor ULLMAN, EDWIN F.SINGH, SHARAT
Owner MONOGRAM BIOSCIENCES
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